Molecular and Cellular Effects of Chemical Chaperone—TUDCA on ER-Stressed NHAC-kn Human Articular Chondrocytes Cultured in Normoxic and Hypoxic Conditions
Abstract
:1. Introduction
2. Results
2.1. Analysis of Cell Viability
2.2. Induction of Hypoxia, Cell Morphology, and Apoptosis
2.3. Analysis of ER Stress
2.4. Analysis of Inflammation
2.5. Analysis of ROS Production
2.6. Analysis of Collagen Production
2.7. Analysis of Membrane Surface Charge Density
3. Discussion
4. Materials and Methods
4.1. Reagents
4.2. Cell Culture and Induction of Hypoxia
4.3. Cell Viability
4.4. Cell Morphological Analysis
4.5. Detection of Apoptosis
4.6. RNA Isolation and Gene Expression Analysis
4.7. Protein Assay
4.8. Sodium Dodecyl Sulfate–Polyacrylamide-Gel Electrophoresis
4.9. Immunoblotting
4.10. Reactive Oxygen Species Generation
4.11. Caspase 1 Activity Assessment
4.12. Zeta Potential Measurements
4.13. Statistical Analysis
5. Conclusions
Author Contributions
Funding
Data Availability Statement
Conflicts of Interest
Sample Availability
References
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Agent | Symbol | Reported Effect | References |
---|---|---|---|
Tauroursodeoxycholic acid | TUDCA | Alleviation of ER stress | [6] |
Alleviation of inflammation | [32] 1 | ||
Alleviation of oxidative stress | [32] 1 | ||
Restoration of collagen expression | [6,33] | ||
Hyaluronic acid | HA | Alleviation of oxidative stress | [34,35,36] |
Alleviation of inflammation | [37,38,39] | ||
Restoration of collagen expression | [40,41] | ||
Interleukin-1β | IL-1β | Induction of inflammation | [5,42,43,44] |
Apoptosis | [21,22,27,42,43,45] | ||
Induction of ER stress | [8,17,22,27] | ||
Induction of oxidative stress | [27,45,46] | ||
Reduction of collagen expression | [21,22,40,41] | ||
Tunicamycin | TNC | Induction of ER stress | [6,11,18,27,47] |
Apoptosis | [6,11,18,47] | ||
Induction of oxidative stress | [48,49] 1 | ||
Reduction of collagen expression | [6,18] |
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Kusaczuk, M.; Naumowicz, M.; Krętowski, R.; Cukierman, B.; Cechowska-Pasko, M. Molecular and Cellular Effects of Chemical Chaperone—TUDCA on ER-Stressed NHAC-kn Human Articular Chondrocytes Cultured in Normoxic and Hypoxic Conditions. Molecules 2021, 26, 878. https://doi.org/10.3390/molecules26040878
Kusaczuk M, Naumowicz M, Krętowski R, Cukierman B, Cechowska-Pasko M. Molecular and Cellular Effects of Chemical Chaperone—TUDCA on ER-Stressed NHAC-kn Human Articular Chondrocytes Cultured in Normoxic and Hypoxic Conditions. Molecules. 2021; 26(4):878. https://doi.org/10.3390/molecules26040878
Chicago/Turabian StyleKusaczuk, Magdalena, Monika Naumowicz, Rafał Krętowski, Bartosz Cukierman, and Marzanna Cechowska-Pasko. 2021. "Molecular and Cellular Effects of Chemical Chaperone—TUDCA on ER-Stressed NHAC-kn Human Articular Chondrocytes Cultured in Normoxic and Hypoxic Conditions" Molecules 26, no. 4: 878. https://doi.org/10.3390/molecules26040878